Principal Investigator/Program Director (Last, first, middle): Mendelsohn, Michael, E RESEARCH &RELATED Other Project Information 1. * Are Human Subjects Involved? m Yes l No 1.a. If YES to Human Subjects Is the IRB review Pending? m Yes m No IRB Approval Date: Exemption Number: 1 2 3 4 5 6 Human Subject Assurance Number 2. * Are Vertebrate Animals Used? l Yes m No 2.a. If YES to Vertebrate Animals Is the IACUC review Pending? l Yes m No IACUC Approval Date: Animal Welfare Assurance Number A3775-01 3. * Is proprietary/privileged information m Yes l No included in the application? 4.a.* Does this project have an actual or potential impact on m Yes l No the environment? 4.b. If yes, please explain: 4.c. If this project has an actual or potential impact on the environment, has an exemption been authorized or an environmental assessment (EA) or environmental impact statement (EIS) been performed? m Yes m No 4.d. If yes, please explain: 5.a.* Does this project involve activities outside the U.S. or l Yes m No partnership with International Collaborators? 5.b. If yes, identify countries: France 5.c. Optional Explanation: 6. * Project Summary/Abstract 2434-abstract.pdf Mime Type: application/pdf 7. * Project Narrative 9229-Project_Narrative.pdf Mime Type: application/pdf 8. Bibliography &References Cited 2013-REFERENCES.HL56069.FINAL.pdMf ime Type: application/pdf 9. Facilities &Other Resources 9502-resources.pg1.pdf Mime Type: application/pdf 10. Equipment Tracking Number: Other Information Page 5 OMB Number: 4040-0001 Expiration Date: 04/30/2008 Principal Investigator/Program Director (Last, first, middle): Mendelsohn, Michael, E The role of hormone action and hormone replacement therapy in vascular biology is an area of intense interest, but remains highly controversial. Most investigators now favor the hypothesis that the evolving nature of vascular disease makes critical the timing of initiation of estrogen therapy, with a protective effect seen when women receive estrogen during or soon after menopause, rather than much later. It is now well accepted that estrogen has direct effects on cardiovascular tissues that account for the majority of the protective effects of estrogen against vascular diseases. The direct effects of estrogen on the vasculature are mediated by two ligand-activated transcription factors, estrogen receptors ER[unreadable] and ER[unreadable]. This competitive renewal application builds upon over a decade of research done by the Molecular Cardiology Research Institute in collaboration with the Vascular Surgery Division at Tufts-New England Medical Center, in which we have explored the mechanisms of ER[unreadable] and ER[unreadable] action in the vasculature using wild type (WT) and ER knockout (ERKO) animals, and a combination of molecular, cellular and vascular models. Work to date has refined the understanding of ER action in the vasculature and supports that ERs mediate both control of vascular tone by estrogen and the protective effects of estrogen on vascular injury and atherosclerosis, in part through regulation of NOS isoforms (eNOS, iNOS) by both genomic and non-genomic pathways. However, studies to date have relied on exploring estrogen effects in mouse models in which an ER is disrupted in the early embryo (whole animal ER knockout mice), so that the ER is deleted from conception in all tissues and cells of the body. The present application proposes to test the mechanistic hypotheses we have evolved over the past decade using exciting new genetically modified mice that are now in hand. These mice, and their tissues and cells, enable the precise deletion of ER[unreadable] or ER[unreadable] from either EC or VSMC and provide powerful approaches to testing the central hypotheses of this application that vascular endothelial cell ER[unreadable] regulates eNOS expression and activity;vascular smooth muscle cell (VSMC) ER[unreadable] mediates estrogen inhibition of VSMC proliferation during vascular injury and atherosclerosis;and VSMC ER[unreadable] controls genes encoding